The human Androgen receptor (AR) is an important target for developing drug therapies combating prostate cancer. The very early event in AR activation, namely the intramolecular interaction between the N- and C-terminus of AR, can now be analyzed with the ChromoTek Fluorescent-Two Hybrid (F2H®) AR: wild-type (wt), as well as several mutant AR forms can be investigated.

Green spot is present, red spot is present at the same location => assay works: interaction

Figure: AR-LBD-GFP (top left) forms bright green spots at the genetic Platform in nuclei of F2H-BHK cells. Before addition of DHT, AR-NTD-RFP (bottom left) is dispersed in the nuclei. Upon 20 min incubation with 10nM DHT, the red spots appear (bottom right, arrows) at the location of the green spots, indicating induction of the interaction between AR-LBD and AR-NTD.

Reversible Screening:

The F2H AR assay is fully reversible: the AR wt interaction can be induced by an agonist (e.g. 0.25nM DHT) and then subsequently disrupted by addition of a potent antagonist (e.g. 10µM bicalutamide).

Movie: Live-cell F2H imaging of reversible interactions between AR-LBD-GFP (green) and AR-NTD-RFP (red) in F2H-BHK cells upon 0.25nM DHT and 10µM bicalutamide treatments. The presence of green spots indicate that AR-LBD-GFP has been anchored to the platform.